title: Brief report: a human induced pluripotent stem cell model of cernunnos deficiency reveals an important role for XLF in the survival of the primitive hematopoietic progenitors.
abstract: Cernunnos (also known as XLF) deficiency syndrome is a rare recessive autosomal disorder caused by mutations in the XLF gene, a key factor involved in the end joining step of DNA during nonhomologous end joining (NHEJ) process. Human patients with XLF mutations display microcephaly, developmental and growth delays, and severe immunodeficiency. While the clinical phenotype of DNA damage disorders, including XLF Syndrome, has been described extensively, the underlying mechanisms of disease onset, are as yet, undefined. We have been able to generate an induced pluripotent stem cell (iPSC) model of XLF deficiency, which accurately replicates the double-strand break repair deficiency observed in XLF patients. XLF patient-specific iPSCs (XLF-iPSC) show typical expression of pluripotency markers, but have altered in vitro differentiation capacity and an inability to generate teratomas comprised of all three germ layers in vivo. Our results demonstrate that XLF-iPSCs possess a weak NHEJ-mediated DNA repair capacity that is incapable of coping with the DNA lesions introduced by physiological stress, normal metabolism, and ionizing radiation. XLF-iPSC lines are capable of hematopoietic differentiation; however, the more primitive subsets of hematopoietic progenitors display increased apoptosis in culture and an inability to repair DNA damage. Together, our findings highlight the importance of NHEJ-mediated-DNA repair in the maintenance of a pristine pool of hematopoietic progenitors during human embryonic development.
authors.affiliation: Institute of Genetic Medicine, Newcastle University, Newcastle, United Kingdom; NESCI, Newcastle University, Newcastle, United Kingdom.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Tilgner
authors.forename: Irina
authors.initials: I
authors.lastname: Neganova
authors.forename: Chatchawan
authors.initials: C
authors.lastname: Singhapol
authors.forename: Gabriele
authors.initials: G
authors.lastname: Saretzki
authors.forename: Jumana Yousuf
authors.initials: JY
authors.lastname: Al-Aama
authors.forename: Jerome
authors.initials: J
authors.lastname: Evans
authors.forename: Vera
authors.initials: V
authors.lastname: Gorbunova
authors.forename: Andrew
authors.initials: A
authors.lastname: Gennery
authors.forename: Stefan
authors.initials: S
authors.lastname: Przyborski
authors.forename: Miodrag
authors.initials: M
authors.lastname: Stojkovic
authors.forename: Lyle
authors.initials: L
authors.lastname: Armstrong
authors.forename: Penny
authors.initials: P
authors.lastname: Jeggo
authors.forename: Majlinda
authors.initials: M
authors.lastname: Lako
mesh_terms: D001483:Base Sequence; D002454:Cell Differentiation; D002460:Cell Line; D002470:Cell Survival; D053903:DNA Breaks, Double-Stranded; D059766:DNA End-Joining Repair; D045643:DNA Repair Enzymes / Q000172:deficiency* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000172:deficiency* / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D008954:Models, Biological; D008969:Molecular Sequence Data
chemical_list: D004268:DNA-Binding Proteins; C508551:NHEJ1 protein, human; D045643:DNA Repair Enzymes
keywords: Cernunnos syndrome; Double-strand break; Human pluripotent stem cells; Induced pluripotent stem cells; Nonhomologous end joining